Crystal structure of the Cys-NO modified YopH tyrosine phosphatase.

Autor: Rocha RF; Laboratório de Biologia Molecular Estrutural, Departamento de Bioquímica, CCB, Universidade Federal de Santa Catarina, Florianópolis 88040-900, Brazil., Martins PGA; Laboratório de Biologia Molecular Estrutural, Departamento de Bioquímica, CCB, Universidade Federal de Santa Catarina, Florianópolis 88040-900, Brazil., D'Muniz Pereira H; São Carlos Institute of Physics, University of São Paulo, São Carlos, Brazil., Brandão-Neto J; Diamond Light Source, Diamond House, Harwell Science and Innovation Campus, Didcot OX110DE, United Kingdom., Thiemann OH; São Carlos Institute of Physics, University of São Paulo, São Carlos, Brazil; Department of Genetics and Evolution, Federal University of São Carlos, São Carlos, Brazil., Terenzi H; Laboratório de Biologia Molecular Estrutural, Departamento de Bioquímica, CCB, Universidade Federal de Santa Catarina, Florianópolis 88040-900, Brazil. Electronic address: hernan.terenzi@ufsc.br., Menegatti ACO; Department of Molecular Biology, Federal University of Paraiba, João Pessoa 58051-900, Brazil. Electronic address: angela.menegatti@dbm.ufpb.br.
Jazyk: angličtina
Zdroj: Biochimica et biophysica acta. Proteins and proteomics [Biochim Biophys Acta Proteins Proteom] 2022 Mar 01; Vol. 1870 (3), pp. 140754. Date of Electronic Publication: 2022 Jan 05.
DOI: 10.1016/j.bbapap.2022.140754
Abstrakt: Protein tyrosine phosphatases (PTPs) are key virulence factors in pathogenic bacteria, consequently, they have become important targets for new approaches against these pathogens, especially in the fight against antibiotic resistance. Among these targets of interest YopH (Yersinia outer protein H) from virulent species of Yersinia is an example. PTPs can be reversibly inhibited by nitric oxide (NO) since the oxidative modification of cysteine residues may influence the protein structure and catalytic activity. We therefore investigated the effects of NO on the structure and enzymatic activity of Yersinia enterocolitica YopH in vitro. Through phosphatase activity assays, we observe that in the presence of NO YopH activity was inhibited by 50%, and that this oxidative modification is partially reversible in the presence of DTT. Furthermore, YopH S-nitrosylation was clearly confirmed by a biotin switch assay, high resolution mass spectrometry (MS) and X-ray crystallography approaches. The crystal structure confirmed the S-nitrosylation of the catalytic cysteine residue, Cys403, while the MS data provide evidence that Cys221 and Cys234 might also be modified by NO. Interestingly, circular dichroism spectroscopy shows that the S-nitrosylation affects secondary structure of wild type YopH, though to a lesser extent on the catalytic cysteine to serine YopH mutant. The data obtained demonstrate that S-nitrosylation inhibits the catalytic activity of YopH, with effects beyond the catalytic cysteine. These findings are helpful for designing effective YopH inhibitors and potential therapeutic strategies to fight this pathogen or others that use similar mechanisms to interfere in the signal transduction pathways of their hosts.
(Copyright © 2022. Published by Elsevier B.V.)
Databáze: MEDLINE
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